Three virulent species of Brucella are currently listed as biological select agent and toxins (BSAT) in the United States. During recent biannual reviews of listed agents and toxins, these virulent...

In many research and diagnostic facilities, liters of biologically contaminated liquid waste are produced on a daily basis and need to be safely disposed of. The gold standard is steam sterilization, namely, in most cases autoclaving for 20 minutes at 121°C. Here we demonstrate the importance of the following parameters when autoclaving liquid waste solutions in a laboratory setting: (1) size of the receptacle for the reference probe, (2) material of the vessel containing the liquid to be autoclaved, and (3) volume of the liquid in the receptacle. If not appropriately controlled, the temperature required to inactivate the liquid might never be reached at all, thus representing a biohazard that should not be underestimated. Our results demonstrate that it is critical to use identical receptacles and liquid volumes for the temperature reference probe and for the liquid to be autoclaved.

The utilisation of the CRISPR/Cas9 technology has sparked a renewed interest in gene drive mechanisms. These mechanisms of biased inheritance may yield promising applications in the fields of vector control and nature conservation. However, the same properties that will enable these applications may also pose a risk if organisms that are equipped with gene drive cassettes are unintentionally released into the environment. Although several groups of scientists and regulators have started to address these safety concerns, there are currently no dedicated guidelines published on the required risk assessment and minimal control measures applicable to gene drive organisms in contained use. To fill this gap, this paper describes a fundamental approach to assessing the risks of these organisms while handled in a contained laboratory environment. Based on the likelihood that an adverse effect will arise from the handling of a gene drive organism and the severity of this effect, three risk classes for contained use activities are presented. Finally, specific minimum requirements regarding physical measures and working practices are proposed according to the presented risk classes and tailored to activities with rodents, insects, and fungi, which are most likely to be used for gene drive applications in the near future.

With increased research efforts on vector-borne diseases such as malaria, dengue, and recently Zika, there has been a need to maintain in captivity arthropods that are known or believed to be involved in the transmission of pathogens to humans in a well-controlled laboratory environment. Therefore, it is pivotal to establish safe yet practical procedures inside an insectary to minimize the associated biosafety risks and prevent the potential escape of the arthropods used into the environment. This article provides a detailed overview of an arthropod disease vectors facility features with a focus on arthropod containment level 2 (ACL-2) laboratory and aims to support health safety and environment (HSE) and biosafety professionals as well as scientists who work on vector-borne diseases by detailing safety procedures inside an ACL-2 environment. In particular, I detail here the associated safety procedures of an ACL-2 and the insectary-specific equipment required for this type of facility. Research on mosquitoes was used here as an example of a disease vector and presents procedures relevant to usage of this vector in the context of Zika virus. The work presented here shows that risks associated with working on infected arthropod vectors can be minimized to an acceptable level and in turn facilitate the research on the diseases they help spread. Similar principles can be applied to other vector-borne diseases research as well, although the work may have a distinct nature.

Applied BiosafetyVol. 22, No. 2 Book ReviewsFree AccessMolecular Microbiology Diagnostic Principles and Procedures, Third EditionReviewed by Rebecca ZaffiniReviewed by Rebecca ZaffiniSearch for more papers by this authorPublished Online:1 Jun 2017AboutSectionsPDF/EPUB Permissions & CitationsPermissionsDownload CitationsTrack CitationsAdd to favorites Back To Publication ShareShare onFacebookTwitterLinked InRedditEmail Molecular Microbiology Diagnostic Principles and Procedures, Third Edition. Editors: David H. Persing, Fred C. Tenover, Randall T. Hayden, Margareta Ieven, Melissa B. Miller, Frederick S. Nolte, Yi-Wei Tang, and Alex van Belkum. Washington, DC: ASM Press, 2016. ISBN: 9781555819088, 810 pp.Molecular diagnostics is a rapidly progressing field, and much has changed since the second edition of Molecular Microbiology was published in 2011. As pressures on clinical laboratories mount to keep up with quality-based care, molecular testing is a must have to provide accurate results in an efficient manner. The third edition of Molecular Microbiology contains an enormous amount of technical information that spans all aspects of microbiology—bacteriology, virology, parasitology, mycology, mycobacteriology, public health, and point-of-care testing.This book is divided into 56 chapters with contributions from many knowledgeable authors. Some chapters from the previous edition have been reorganized to stay relevant with the current testing landscape. New sections include Molecular Diagnostics and Public Health, Syndromic Diagnostics, and Developing a Business Case for Hospital Administration.A section that was particularly well put together includes Section I on Novel and Emerging Technologies. New technologies can be a challenge for a clinical laboratory to implement into everyday practice. This section well described validation approaches for technologies such as next-generation sequencing and matrix-assisted laser desorption ionization–time of flight and covered topics spanning from sample preparation to appropriate resulting. In this same vein, I found that Section V on Syndromic Diagnostics could have included better recommendations for validations of multiplex assays.As laboratory testing shifts from traditional methods to molecular methods, the laboratory is shifting from its traditional business role in an institution to a cost center. This puts additional stress on the microbiology laboratory to prove its value to the institution. Developing a business case for new, expensive testing is a new challenge for the clinical laboratory. Chapter 55 discusses this topic well, although I would suggest a full example be included and more resources to help guide laboratory managers with their own cases.The intended audience for this textbook is the clinical laboratory technologist, manager, and medical director. The great benefit in this book is the multitude of easy-to-read tables outlining different testing options currently available and their strengths and weaknesses. For example, Tables 1 and 2 on pages 173-175 are incredibly informative and nicely summarize commercial options available for methicillin resistant S.aureus/methicillin susceptible S.aureus (MRSA/MSSA) molecular testing. Another nicely organized table is Table 1 on pages 341-342 that describes commercially available molecular systems for bloodstream infections. With an extensive array of options now on the market, the clinical laboratory gets overwhelmed with comparing platform and assay pros and cons and likely has little time to do a full investigation; this book provides a great tool as a place to start considering possible testing selections. This text is also an informative resource for infectious disease physicians and infection preventionists to better understand the utility of tests currently in use at their institutions. I highly recommend this text as a resource for the clinical microbiology laboratory and infectious disease departments.FiguresReferencesRelatedDetails Volume 22Issue 2Jun 2017 Information© ABSA International 2017To cite this article:Reviewed by Rebecca Zaffini.Molecular Microbiology Diagnostic Principles and Procedures, Third Edition.Applied Biosafety.Jun 2017.75-76.http://doi.org/10.1177/1535676017701168Published in Volume: 22 Issue 2: June 1, 2017PDF download

Models that are simple to use and interpret and that predict aerosol concentration and integrated exposure dose are needed for local, timely, and cost-effective evaluation of risks associated with biocontainment operations. A material balance model is presented with explicit assumptions that provide a safety context for assessing potential laboratory operating and accident scenarios (1) to support determination of aerosol concentration when limited experimental data are available or (2) to assist in the interpretation of experimental results. The model is used to assess recent experimental data for large animal primary containment systems in animal biosafety level 4; it is also applied to estimate integrated dose from particle concentration for assessments of potential exposure of laboratory workers. The model is further shown to reproduce previously published laboratory accident results.

One of the most critical aspects of safe operations in biosafety level 3 and 4 and animal biosafety level 3 and 4 biocontainment laboratories is the effective inactivation of biological select agen...

In December 2014, the National Academies of Sciences convened the first of 2 meetings to discuss the potential risks and benefits of so-called gain-of-function (GOF) research, which was defined by the National Scientific Advisory Board for Biosecurity as research toward increasing the transmissibility, pathogenicity, or ability to evade countermeasures of pathogens with pandemic potential. The symposium was part of a deliberative process undertaken by the United States (US) government to inform in developing guidelines for the safe conduct of GOF research. Participants discussed scientific, technical, and ethical aspects regarding the conduct of research, biosafety measures in the US, and, to a minimal degree, biosafety in other countries. The dearth of information available regarding codified international biosafety requirements and norms, as well as the stringency and verification of their implementation, is particularly troubling as many countries conduct or plan to conduct GOF research. This analysis was conducted to inform policy makers, scientists, and biosafety professionals on a global scale of strengths and potential safety weaknesses by comparing publicly available regulations, laws, guidelines, and published articles describing biosafety processes and measures in other developed countries that may conduct GOF research compared to measures in place in the US. As GOF research has the potential to cause pandemic illness, the findings in this review can be applied to address potential safety gaps by countries with established biosafety capabilities and can serve as a comprehensive guide for countries with nascent or developing biosafety capability working with high-consequence pathogens or contemplating conducting GOF research.

Applied BiosafetyVol. 21, No. 3 Special FeaturesFree AccessCapsuleRobert A. RasmussenRobert A. RasmussenCorresponding Author: Robert A. Rasmussen, Wyss Institute for Biologically Inspired Engineering, 3 Blackfan Circle, Boston, MA 02215, USA. E-mail Address: robert.rasmussen@wyss.harvard.eduWyss Institute for Biologically Inspired Engineering, Boston, MA, USASearch for more papers by this authorPublished Online:1 Sep 2016AboutSectionsPDF/EPUB Permissions & CitationsPermissionsDownload CitationsTrack CitationsAdd to favorites Back To Publication ShareShare onFacebookTwitterLinked InRedditEmail What’s new, what’s hot, what’s timely? If you don’t have time to search the Internet for the latest developments that might affect your work environment, you just might find some of that information in this Capsule column. Please email any comments, suggestions, or insights to Robert Rasmussen at robert.rasmussen@wyss.harvard.edu.Therapeutic Approaches to RNA Viruses That Exploit Their High Mutation RatesDue to their reliance on error-prone viral RNA polymerases for replication, RNA viruses exhibit very high rates of genetic mutation. Combined with selective pressure, these high mutation rates have imposed enormous difficulties in designing clinical treatments, as well as vaccines, against pathogenic RNA viruses because of the rapid emergence of drug-resistant and/or neutralizing antibody-resistant strains of RNA viruses. A recent review by Tanner et al1 describes 2 therapeutic approaches that in theory could be efficacious despite high rates of viral mutation. Neither approach is aimed at viral enzymes or receptors, the current targets for most antiretroviral therapies. Instead, they are aimed, either directly or indirectly, at structural viral components.The first approach is called dominant drug targeting through dominant-negative interactions, and the authors use the biology of viral capsid synthesis to demonstrate how the approach would operate. Viral capsids are polymerized intracellularly from whatever oligomeric precursors are available. When exposed to a drug that targets viral capsid precursors, selective pressure plus the continuous generation of viral RNA genome mutations can result in the formation of drug-resistant capsid precursor proteins. The result can be chimeric capsids composed of oligomeric precursor proteins encoded by both drug-sensitive and emergent drug-resistant genomes. Dominant-negative interactions occur during the assembling of both drug-resistant and drug-sensitive precursors into chimeric capsids because even if the capsid contains only a small minority of drug-sensitive proteins, the entire capsid and virus particle still remains drug sensitive. Proof-of-concept experiments for dominant drug targeting have already been successfully performed for both poliovirus and dengue virus infections.The second approach is based on exploiting the natural process of viral genetic interference. In any RNA virus-infected cell, error-prone RNA polymerase activity results in an intracellular mix of divergent viral RNA molecules. Genetic interference of the optimal replication kinetics of wild-type viruses occurs due to competition between wild-type and mutant viral genomes for a limited number of viral-packaging proteins as part of the encapsidation mechanism. To exploit genetic interference for treating RNA virus infections, a treatment approach would be to engineer so-called therapeutic infectious particles, or TIPs, containing interfering RNA to out-compete wild-type viruses. To have a competitive advantage, the number of intracellular genomic TIP RNA molecules must exceed the number of wild-type viral RNA molecules. By limiting the size of TIP RNA to encode only the necessary elements for efficient TIP genome replication and packaging, the length of TIP RNA is much shorter than wild-type viral genome RNA. For reasons that have to do either directly with their shorter length or with their higher affinity for certain RNA polymerases, shorter RNA genomes have a replication advantage over longer RNA genomes, thus ensuring that the number of TIP RNA molecules exceeds that of wild-type virus. This gives the relatively abundant TIP RNA genome a competitive stoichiometric advantage over the full-length RNA viral genome for being packaged, or “mobilized,” into the available virus-encoded capsids. As described using human immunodeficiency virus 1 (HIV-1) infection as a prototype for this approach, the authors suggest employing a lentiviral TIP to parasitize HIV-1 infected cells, with the lentiviral TIP genome essentially using HIV-1 as a packaging virus. Essentially, this controlled genetic interference via TIPS would keep replication of the original infecting RNA virus replication at low levels.Although using a lentiviral vector against HIV has been previously demonstrated (eg, using a lentiviral vector encoding a small hairpin RNA against HIV-1 env), the lentiviral vector was eventually lost over time. In contrast, the parasitic nature of using lentiviral TIPs against HIV-1 as described here would ensure that the therapeutic infectious particles remain as long as they are needed—that is, as long as there is HIV packaging. While no experimental data were shown using a potential TIP, in silico model analysis indicated the approach would be efficacious, and epidemiological models predict population infection prevalence would drop.Reference 1. Tanner E Kirkegaard K Weinberger L . Exploiting genetic interference for antiviral therapy. PLoS Genet. 2016;12(5):e1005986. Crossref, Medline, Google ScholarFiguresReferencesRelatedDetails Volume 21Issue 3Sep 2016 Information© ABSA International 2016To cite this article:Robert A. Rasmussen.Capsule.Applied Biosafety.Sep 2016.156-157.http://doi.org/10.1177/1535676016661773Published in Volume: 21 Issue 3: September 1, 2016PDF download